Fig. 1

Abnormal upregulation of NFAT1 expression enhances the proliferation of renal cancer cells. A Determination of the NFAT1 mRNA expression level by the GEPIA web tool. Boxplot analysis of the expression level showing log2 (TPM + 1) on a log-scale. The NFAT1 expression was compared between ccRCC patient specimens and nontumor renal tissues. B and C. Western blot (B) and qRT-PCR analysis (C) of the protein and mRNA expression of NFAT1 in 12 paired renal cancer tissues (T) and the matched adjacent normal tissues (ANT) of the same patient. GAPDH served as an internal reference. ***, P < 0.001. The difference was compared between ANT group and tumor tissue group. D Determination of the overall survival NFAT1 mRNA expression level by the Human Protein Atlas database (P < 0.001) of RCC patients. The difference was compared between low NFAT1 expression and high NFAT1 expression group. E Determination of the biological role of NFAT1 by the CancerSea cancer single-cell state atlas (http://biocc.hrbmu.edu.cn/CancerSEA/) in RCC patients. F and G. Western blot (F) and qRT-PCR (G) analysis of NFAT1 expression in renal cancer cells infected with shControl or shNFAT1s. GAPDH served as an internal reference. For qRT-PCR analysis, data presented as the mean ± SD of three independent experiments. ***, P < 0.001. The differences were compared between shControl group and shNFAT1 group. H and I. Renal cancer cells infected with shControl or shNFAT1s were harvested for MTS (H) and colony formation assay assays (I). Each bar represents the mean ± SD of three independent experiments. **, P < 0.01 ***, P < 0.001. The differences were compared between shControl group and shNFAT1 group. J 786-O cells infected with or without shNFAT1 and/or Tsin-NFAT1 were harvested for MTS assay. ***, P < 0.001. The difference was compared between shControl+Tsin-EV and shNFAT1 + Tsin-EV group, or shNFAT1 + Tsin-EV and shNFAT1 + Tsin-NFAT1. K-M. 786-O cells infected with or without shNFAT1 and/or Tsin-NFAT1 were subcutaneously injected into nude mice. The tumors were harvested and photographed (K) on day 23. Data for tumor mass (L) and tumor volume (M) are shown as the mean ± SD (n = 6). ***, P < 0.001. The difference was compared between shControl+Tsin-EV and shNFAT1 + Tsin-EV group, or shNFAT1 + Tsin-EV and shNFAT1 + Tsin-NFAT1.